US4735907A — Stabilized fluorescent rare earth labels and labe…

Claim 1

Independent Claim 121 words

A fluorescent labeled physiologically reactive species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 2

Dependent Claim 121 words

A fluorescent labeled physiologically reactive species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 3

Dependent Claim 16 words

The labeled species of claim 1 wherein said physiologically reactive species is an antigen or antibody.

Claim 4

Dependent Claim 28 words

The labeled species of claim 1, wherein said chelate is a chelate of europium or terbium and a 1, 3-diketone, phthalate, naphthoate, dipyridine, terpyridine or phenanthroline chelating agent.

Claim 5

Dependent Claim 19 words

The labeled species of claim 1, wherein said labeled physiologically reactive species is a labeled specific binding ligand analog.

Claim 6

Dependent Claim 87 words

The labeled species of claim 1, wherein said polymer comprises: (a) from about 75 to about 92 weight percent of recurring units derived from styrene or a water-insoluble substituted styrene derivative, (b) from about 5 to about 15 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer containing a hydroxy or amide group, and (c) from about 3 to about 10 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 7

Dependent Claim 88 words

The labeled species of claim 5 wherein said rare earth chelate is a chelate of europium, said physiologically reactive species is a hapten, and said polymer comprises: (a) from about 75 to about 92 weight percent of recurring units derived from styrene or vinyltoluene, (b) from about 5 to about 15 weight percent of recurring units derived from acrylamide, methacrylamide, N-isopropylacrylamide, 2-hydroxyethyl acrylate or 2-hydroxyethyl methacrylate, and (c) from about 3 to about 10 weight percent of recurring units derived from methacrylic acid, acrylic acid or itaconic acid.

Claim 8

Dependent Claim 140 words

A dry analytical element useful in diagnostic or analytical methods comprising an absorbent carrier material containing a fluorescent labeled physiologically reactive species, said labeled species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 9

Dependent Claim 140 words

A dry analytical element useful in diagnostic or analytical methods comprising an absorbent carrier material containing a fluorescent labeled physiologically reactive species, said labeled species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 10

Dependent Claim 169 words

A dry analytical element useful for the determination of a specific binding ligand in an immunoassay, said element comprising a support having thereon one or more zones, one of which is a porous spreading zone, and in one or more of said zones, a fluorescent labeled specific binding ligand analog of said ligand, said labeled analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 11

Dependent Claim 169 words

A dry analytical element useful for the determination of a specific binding ligand in an immunoassay, said element comprising a support having thereon one or more zones, one of which is a porous spreading zone, and in one or more of said zones, a fluorescent labeled specific binding ligand analog of said ligand, said labeled analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 12

Dependent Claim 49 words

The element of claim 8 wherein said rare earth metal is europium or terbium, and said polymer comprises from about 75 to about 92 weight percent of (a), from about 5 to about 15 weight percent of (b), and from about 3 to about 10 weight percent of (c).

Claim 13

Dependent Claim 11 words

The element of claim 8, wherein said ligand is a hapten.

Claim 14

Dependent Claim 33 words

The element of claim 8 further comprising a receptor for said ligand which is immobilized in said element in a manner that keeps it isolated from said ligand analog prior to an assay.

Claim 15

Independent Claim 190 words

A method for the determination of a specific binding ligand in an aqueous liquid, said method comprising the steps of; A. in the presence of a receptor for said ligand, contacting a sample of said liquid with a fluorescent labeled specific binding ligand analog, said ligand analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group to form a complex between said receptor and said ligand analog, B. separating said complex from uncomplexed ligand analog, and C. fluorometrically detecting either said complexed or uncomplexed ligand analog.

Claim 16

Independent Claim 190 words

A method for the determination of a specific binding ligand in an aqueous liquid, said method comprising the steps of; A. in the presence of a receptor for said ligand, contacting a sample of said liquid with a fluorescent labeled specific binding ligand analog, said ligand analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group to form a complex between said receptor and said ligand analog, B. separating said complex from uncomplexed ligand analog, and C. fluorometrically detecting either said complexed or uncomplexed ligand analog.

Claim 17

Dependent Claim 11 words

The method of claim 12 wherein said ligand is a hapten.

Claim 18

Dependent Claim 12 words

The method of claim 12 wherein said complexed ligand analog is detected.

Claim 19

Dependent Claim 143 words

A diagnostic test kit for the determination of a specific binding ligand, said kit comprising (i) a receptor for said ligand, and (ii) a fluorescent labeled immunologically reactive ligand analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 20

Dependent Claim 143 words

A diagnostic test kit for the determination of a specific binding ligand, said kit comprising (i) a receptor for said ligand, and (ii) a fluorescent labeled immunologically reactive ligand analog comprising said ligand bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from a water-insoluble ethylenically unsaturated polymerizable monomer, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 21

Dependent Claim 145 words

A fluorescent labeled physiologically reactive species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from one or more water-insoluble ethylenically unsaturated polymerizable monomer selected from the group consisting of styrene, α-methylstyrene, p-bromostyrene, vinyltoluene, 1-vinylnaphthalene, methyl acrylate, methyl methacrylate, ethyl methacrylate, n-butyl acrylate and propyl acrylate, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Claim 22

Dependent Claim 145 words

A fluorescent labeled physiologically reactive species comprising a physiologically reactive species bound to a fluorescent label comprising a fluorescent rare earth chelate incorporated into a polymeric particle which is derived from a loadable latex having a discontinuous phase and an aqueous phase, said discontinuous phase consisting essentially of a polymer comprising: (a) from about 50 to about 96 weight percent of recurring units derived from one or more water-insoluble ethylenically unsaturated polymerizable monomer selected from the group consisting of styrene, α-methylstyrene, p-bromostyrene, vinyltoluene, 1-vinylnaphthalene, methyl acrylate, methyl methacrylate, ethyl methacrylate, n-butyl acrylate and propyl acrylate, (b) from about 2 to about 30 weight percent of recurring units derived from a nonionic water-soluble or water-dispersible ethylenically unsaturated polymerizable monomer, and (c) from about 2 to about 20 weight percent of recurring units derived from an anionic ethylenically unsaturated polymerizable monomer containing at least one carboxy group.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1991-04-05	1991-10-05	1992-04-05	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1995-04-05	1995-10-05	1996-04-05	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1999-04-05	1999-10-05	2000-04-05	$7700.00	Window Closed / Lapsed

Stabilized fluorescent rare earth labels and labeled physiologically reactive species

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